Cryostorage method and device

ABSTRACT

With a method for cryo-preservation, at least one specimen is arranged on a storage substrate and specimen data, which are characteristic for features of the specimen, are stored at specific positions. Also, a storage substrate for cryo-preservation with such a method is described.

[0001] The present invention relates to methods for cryo-storage ofspecimens, in particular, for the production, storage, and manipulationof biological specimens in a cryo-preserved or in a thawed condition,such as, for example, a cryo-preservation method for biological cells.The invention relates also to methods for writing and reading of data.The invention further relates to a device for cryo-storage of specimens,in particular, a storage substrate for biological specimens, such as,for example, cells or cell components, a device for writing and readingof data in storage media and a cryo-bank system. The invention alsorelates to uses of the cryo-preservation of biological specimens.

[0002] Cryo-preservation is a commonly known method for maintaining inparticular biologically or medically relevant materials. These materialsinclude, for example, tissue and organs, body fluids, or also individualcells or cell components. The cryo-preservation takes place according topredetermined procedures in containers or on substrates, whose shape isadapted to the material or specimen. Containers for cryo-preservationare known, for example, for tissue and organs (see DE-OS 199 22 31, EP-A0 853 238, DE-OS 197 25 768, DE-OS 199 05 163), for blood components(see, for example, DE-OS 198 26 350), and for cell or drop-shapedcryo-specimens (see, for example, U.S. Pat. No. 5,275,016, EP-B 0 475409, DE-OS 199 21 236, EPB 0 804 073).

[0003] A general concern with the cryo-preservation of biologicalspecimens is in the ability of the specimen to be identified.Cryo-preservation specimens must be able to be identified with referenceto their origin and characteristics with a high degree of certainty,without the necessity of thawing. With the macroscopic specimens, thisis not a problem, since organ or blood containers can be provided withan inscription. Locating the cryo-specimen takes place in dependence onstorage systems of the respective cryo-bank.

[0004] With small cryo-specimens in the form of frozen suspension drops,cells, cell aggregates, or cell components, the identification of thecryo-specimens is considerably problematic. A cryo-specimen would benegligibly small compared with the inscription. Often, an interestexists in the cryo-preservation of a plurality of microscopically smallspecimens. The storing and identification of small cryo-specimens withinscriptions would be impractical. In addition, the cryo-preserved cellspecimens are available in a disordered state with common preservationtechniques, which are based on the spraying of cell suspensions on coolsurfaces (see, for example, EP-B 0 475 409). Only large amounts ofindividual specimens can be preserved commonly and unspecifically.

[0005] With the preservation techniques described in EP-B-8-4 073 andDE-OS 199 21 236, an arranged placement and specific processing of eventhe smallest specimens on the cryo-substrates is possible. The specimendeposition takes place, for example, with the use of a micro-dropshooting device, which is controlled on the basis of predeterminedtarget coordinates. The specimens are located on defined substratepositions, on which also a specific measurement of specimen propertiesand identification of the specimens is possible. The substrate can beprovided with a marking, in order to define the specimen positions onthe substrate. For example, in DE-OS 199 21 236, for the matrix-typedeposition of cryo-specimens in straight lines and columns, it isproposed that the substrate is provide with a designation of the columnsand lines. This marking technique is illustrated in FIG. 27 (prior art).

[0006] The conventional marking of cry-substrates according to FIG. 27has the following disadvantages. While the identification of specimensis possible, however, only information about the positions is provided.The limited information content of the substrate marking, however,represents a problem since, in addition to the specimen identification,also additional data, for example, about the condition or the history ofthe specimen or about measurement results should be available. Thesedata can be stored in a parallel-operated data bank. The separateoperation of cryo- and data banks, however, represents a considerablerisk for the certainty of the feature allocation to the individualspecimens. This risk is critical in particular with human medical uses,since a specific mistake can defeat the success of a further use of thecryo-specimen. In addition, the substrate marking has the disadvantagethat specimen identification is possible only in connection with thecryo-substrate. When a specimen removal takes place, for example, suchas that described in DE-OS 199 21 236, after separation from thecryo-substrate, a specimen identification can take place only by anexpensive measurement of specific characteristics in a thawed state.

[0007] From DE-OS 197 52 085, a specimen carrier for microscopicanalysis of a plurality of specimens is known. The conventional specimencarrier is formed as a substrate with a plurality of specimen receivingspaces, as shown in FIG. 28 in schematic plan view (prior art). Thesubstrate, for example, has the shape of a plate storage medium (forexample, a CD). Between a passage hole in the substrate center and theassociated matrix-type specimen-receiving spaces, a ring region isformed. From DE-OS 197 52 085, it is known to form these ring regionsfor storing specimen data. The common specimen carrier has thedisadvantage that it can only be used for receiving liquid specimens andnot for cryo-preservation. In addition, the storage of specimen data onthe inner ring represents the same disadvantage as the above-describedsubstrate marking. More data can be stored; however, the association tothe individual specimens is not possible without errors.

[0008] In addition to the noted disadvantages of the common technology,the following basis also exists for the minimally developed use, untilnow, of cryo-preservation, in particular, in cellular biotechnology. Ifa direct freezing of biological specimens takes place in a liquidcooling phase (for example, nitrogen), a risk of contamination exists.Over the cooling phase, viruses can be transmitted to the specimens. Inorder to avoid this risk, the contact with the liquid phase must beavoided or a sealed covering of the specimens must take place. Up tonow, this has not been able to be realized in a practical manner.

[0009] From labor technology, specimen carriers, for example, in theform of object carriers or micro-titer plates are known, which areequipped with data storage media. These conventional specimen carriersare not suited for cryo-storage. First, they are merely suited for a useat ambient room temperature or a refrigerator temperature above thefreezing point of water. A use at low temperatures has not been providedup to this point. Second, conventional specimen carriers are provided assubstrates for specimens. Treatment, manipulation, or cultivation of thespecimens takes place on the substrates. For storage in a preservedstate, the conventional specimen carriers, however, are not suitable. Inthis connection, the containers for cryo-preservation are used, asdescribed above. Finally, the conventional specimen carriers are notsuitable for an effective specimen storage and manipulation. Inpractice, they must be manually transported; storage with high densityis impossible.

[0010] The object of the present invention is to provide an improvedmethod for cryo-preservation, with which the disadvantages of theconventional techniques are overcome, which has a broader range of use,and in particular, which is suited for automated preservation storage.The new method for cryo-preservation should make possible, inparticular, that the specimen data are accommodated in greater amountsand with a higher data certainty (that is, with increased certainty ofthe association of specimen data to determined specimens). The inventionalso makes possible a highly specific data association to individualcryo-specimens. The object of the invention is also to provide devicesfor implementation of the improved cryo-preservation methods describedabove.

[0011] These objects are solved with the methods or devices with thefeatures of patent claims 1, 12, 21 and 22. Advantageous embodiments anduses of the invention are provided in the dependent claims.

[0012] The basic idea of the present invention is to arrange at leastone specimen on a substrate and on the substrate, to provide storage ofspecimen data, which are characteristic for the features of thecryo-specimen. The storage of the specimen data takes placeposition-specific in a specimen data storage medium, preferably, at thestorage position of the respective specimen. By means of the storage ofspecimens and specimen data on common or closely adjacent or adjoiningpositions of the substrate, a series of advantages is achieved. Thespecimen data are clearly associated by means of their storage positionsto the respective specimens. A mistake in specimen association isimpossible. Upon removal of specimens, the associated data can besimultaneously read or removed with the storage medium from thesubstrate, so that also, after the removal of the specimen, with furtherprocessing, the identification of the specimen and the association ofthe specimen data are ensured. A removal of individual specimens atdesired temperatures, in particular, also in a frozen state, can takeplace.

[0013] A considerable improvement that can be achieved with the presentinvention is that, first, methods and suitable devices can be provided,which are directly optimal for storage and preservation of biologicalspecimens over long periods of time (months and years) at lowtemperatures (for example, below −50° C.). With the invention, a newarea of use with operating temperatures for the use of data storage hasbeen made available, which, prior to the invention, was not utilized.

[0014] Depending on the use, the specimen deposition and the datastorage can take place at ambient room temperature with subsequentcooling to a required preservation temperature or also in a cooledstate. The inventors have surprisingly found that writing as well asreading of data in or out of the known storage media (for example,optical storage, magnetic storage, electromagnetic storage, FLASHstorage), or in special storage media for the object of cryo-storage atpreservation temperatures beneath the freezing point of water arepossible. The specimen data sets are reliably readable in all phases ofa cryo-preservation process. The readability of data storage at thesetypes of low temperatures, that likewise, exclude a writing of data,represents an advantage known to the inventors, which amounts to abroader usability of the inventive cryo-storage.

[0015] Particularly advantageous to the present invention is thedeposition of a plurality of specimens on a common storage substratewith a position-specific storage of a plurality of specimen data sets.The storage substrate serves simultaneously as a cryo-substrate withspecimen carriers for receiving, retaining, and releasing cryo-specimensand as data carriers, which stores a plurality of data on the substrateposition corresponding to the respective specimen positions, as in astorage medium known in computer technology. The specimens are appliedin portions (for example, in drops) and isolated from one another ascell suspension volumes (for example, as cell suspension drops) on or inthe specimen carrier of the cryo-substrate. Each specimen carrier isassociated with a specimen data storage medium, in which associated dataare stored. The simultaneous storage of specimens and specimen datatakes place, such that it is stable over long periods of time and it issafeguarded from mix-ups. The inventive cryo-preservation is designatedas “cryo-storage”, because of the analogy to electronic data storage.

[0016] According to a preferred embodiment of the invention, thecryo-storage takes place on a storage substrate with at least onecryo-storage element. Each cryo-storage element includes a specimencarrier and a specimen data storage, which form therewith an integralcomponent, which is removable reversibly or irreversibly from thesubstrate. The specimen carrier and the data storage form a secureconnection, which is attached removably on the storage substrate forcryo-storage. In order to remove a specimen from the storage substrate,the entire cryo-storage element is removed from the storage substrate. Asubject of the present invention is also the cryo-storage element, whichincludes a specimen carrier for receiving a cryo-specimen and a specimendata storage for storage of associated specimen data. According to apreferred embodiment of the invention, a storage substrate is formed bya base body, which preferably has a flat shape, with a plurality ofcryo-storage elements. The storage substrate can have a predeterminedtwo- or three-dimensional geometric form, depending on the use.According to a preferred embodiment of the invention, the base body ofthe storage substrate has the form of an optical storage plate (CD-ROM),in which the cryo-storage elements are integrated, or at least onecircuit board, on which the cryo-storage elements, like electricalcircuits (chips), are placed.

[0017] A subject of the present invention is also a method for operatinga cryo-bank with a plurality of storage substrates. On at least onestorage substrate, a plurality of specimens are stored, which, forexample, belong to an organism (test subject). The specimens include,for example, one or more specific cells of the test subject (forexample, stem cells, tissue cells). First, the storage of the specimenstakes place commonly with the specimen-specific data, in particular,data for identification of the type of specimen and test subject,identification of the preservation time point, and identification ofmeasurement data at the time point of the preservation. In the operationof the cryo-bank, specimens are removed commonly with the associatedspecimen data for measurement purposes, diagnostic objectives, ortherapeutic methods, and/or to supplement further specimens or specimendata. The specimen data include generally all features and parameters ofthe specimen and the specimen donor, and if need be, additionalinformation for data storage on the storage substrate. An inventivelyused specimen data storage has a storage capacity of at least 4megabytes, for example.

[0018] By means of the following advantageous features of the inventivecryo-storage, the disadvantages of conventional planar orthree-dimensional substrates are overcome. The specimens (for example,frozen cell suspension volumes) are specifically accessible at any pointin time. This is also true for a low temperature state. The smallestspecimen volumes can be arranged at defined substrate positions, whichhave characteristic dimensions in the mm-range or less, preferably,however, typically amounts of 10³ μm³ (10·10·10 μm³) to some 10 nm³. Thespecimens can contain, for example, one or a plurality of cells (10⁵ to10⁶ cells), cell components, biologically relevant objects (such as, forexample, membrane vesicles, DNA materials, macromolecules) and/or cellbonds. The specimens can be arranged with a high density, thecryo-storage having an increased effectiveness.

[0019] The specimens can be removed selectively in a deep-cooled stateof the storage substrate, without interrupting the cooling of theremaining specimens. For specimen reduction and data reading, no thawingof the entire storage substrate is necessary.

[0020] The specimen data can be automatically written or read supportedby a computer. The association of the specimen data storage and specimencarriers is unique. The specimens are stored, such that they aresafeguarded from misidentification. The association of the removedspecimens to the specimen data and also to the storage substrate ismaintained, so that the history of a specimen can be recapitulated. Thisrepresents a particular advantage with medical uses of the invention.

[0021] The cryo-storage elements of the present invention are preferablymade with the use of low-temperature compatible plastic material, which,on the one hand, forms the specimen carrier and on the other hand, formsan embedding for the specimen data storage. The plastic material cantolerate repetitious temperature changes without change and withoutdamage. Preferably, a plastic material is used, whose water absorptioncapacity amounts to <1% of the dead weight, in particular, <0.1% of thedead weight. The cryo-storage elements according to the presentinvention are based, for example, on polyurethane,poly-tetra-fluoroethylene, or polyethylene. The storage substrate of thepresent invention advantageously has a high mechanical stability and along-term durability. The inventive cryo-storage makes possible for thefirst time a secure storage of biological specimens over decades. Acryo-bank can be reliably operated over the entire longevity of a donor(test subject), for example, for the duration of a human life. Thestorage substrate has a relatively simple structure, which makespossible an abundant use of the storage substrates in cryo-banks.

[0022] The invention also has advantages with reference to the notedcontamination risks. The inventive storage substrate makes possible aseries of features to be described below, by means of which a liquidcooling medium is prevented from coming into contact with the specimens.Viral contamination is avoided over the cooling phase. Also, thecondensation of water or other substances on the specimens isimpossible.

[0023] A further important advantage of the storage substrate of thepresent invention is that the stored specimens are accessible in acryo-preserved or thawed state of the common measurement and analysismethods (for example, optical measurement, microscopic analyses) withsimultaneous readability of the data phases. The data in the specimendata storages are also maintained with multiple freezing or thawingprocesses.

[0024] The cryo-storage takes place with conservation conditions chosendepending on the application. The temperature of the cryo-storage andthe chronological cycle of decreases or increases in temperature areselected based on the preservation object and the material. Thepreservation temperature lies in a region below the ambient roomtemperature, preferably, below the freezing point of water with normalpressure and with the preferred long-term use, below −80° C. Thecryo-temperature is preferably adjusted by means of a liquid coolingmedium (nitrogen) or the vapor of the cooling medium.

[0025] The invention has the advantage that the smallest specimenvolumes can be cryo-preserved. This makes possible fast temperaturechanges, reproducible preservation conditions, and an individualspecimen manipulation, treatment, or measurement.

[0026] Further advantages and characteristics of the invention aredescribed with reference to the attached drawings. In the drawings,

[0027]FIG. 1 is a schematic sectional view of a part of the inventivestorage substrate according to a first embodiment of the invention;

[0028]FIG. 2 is a schematic illustration of a specimen removal from astorage substrate according to FIG. 1;

[0029]FIGS. 3 through 7 are schematic plan views of various forms ofspecimen and storage arrangements on a storage substrate;

[0030]FIG. 8 is a schematic sectional view of a part of a storagesubstrate according to a second embodiment of the invention;

[0031]FIG. 9 is a schematic view of various geometric arrangements ofcryo-storage elements;

[0032]FIG. 10 is a schematic sectional view of a part of a storagesubstrate according to a third embodiment of the invention;

[0033]FIG. 11 is an illustration of the removal of cryo-specificelements from an inventive storage substrate according to a fourthembodiment of the invention;

[0034]FIG. 12 is a schematic illustration of a film-like cover on aninventive storage substrate;

[0035]FIG. 13 is a schematic sectional view of a part of a storagesubstrate according to a fifth embodiment of the invention;

[0036]FIG. 14 are schematic perspective views of storage substratesaccording to a sixth embodiment of the invention;

[0037]FIG. 15 is a schematic perspective view of a cryo-storage elementof the storage substrate according to FIG. 14;

[0038]FIG. 16 is a schematic perspective view of a modification of thecryo-storage element according to FIG. 15;

[0039]FIGS. 17 through 20 are schematic perspective views of theinventive cryo-storage element according to further embodiments of theinvention;

[0040]FIGS. 21 and 22 are schematic illustrations of the loading of thespecimen carriers and the removal of the specimens;

[0041]FIGS. 23 through 26 are schematic perspective views of theinventive storage substrate according to further embodiments of theinvention; and

[0042]FIGS. 27 and 28 are plan views of conventional specimen carriers(prior art).

[0043] According to the present invention, at least one specimen isarranged in a specimen carrier on a substrate and on the substrate,storage of specimen data is made, which are characteristic for featuresof the cryo-specimen. The storage of the specimen data takes placeposition-specific in a specimen data storage, preferably, at the storageposition of the respective specimen. The connection on a specimencarrier for receiving a cryo-specimen and a specimen data storage forstorage of associated storage data is designated as a “cryo-storageelement”. An inventive storage substrate is preferably formed as a basebody, which preferably, has a flat form, with a plurality ofcryo-storage elements.

[0044] With the first embodiment of the inventive storage substrate 100shown in FIG. 1, a base body 110 is provided, which supports a pluralityof cryo-storage elements 120. The base body 110 (shown in cut-awaymanner) has typical dimensions, such as, for example, an opticallyreadable/writable storage plate (in the following: CD, diameterapproximately 12 cm, for example). For receiving the cryo-storageelement 120, the base body 110 has passage openings 111, in which thecryo-storage elements sit in a type of a press fit. On one side of thebase body 110, a film-type storage medium 112 is arranged. The storagemedium 112 is a data layer, such as the type that is known for commonCD's and that is suitable for reading and writing of data. The storagemedium 112 preferably is designed for optical writing (“burning”) andreading of data. However, it also can be a magnetic or a topographicstorage medium. On the storage medium 112, a protective layer isprovided, if necessary (not shown).

[0045] The storage medium 112 includes layer regions, which lie on thebase body 110 and serve as base storage 113, and regions, which areassociated with the cryo-storage elements 120 and serve as specimen datastorage 122. The base storage 113 and specimen data storage 122 firstcan form a closed layer of the storage medium 112, which after specimenremoval (see FIG. 2) is broken, when necessary. The base storage 113contains preferably substrate data, which, for example, relate to thetype of the arrangement of the cryo-storage elements and theidentification of the substrates. The specimen data storage 122 containsspecimen data (see below).

[0046] The cryo-storage elements 120 comprise, respectively, a specimencarrier 121 and the specimen data storage 122. The specimen carrier 121is a formed component made of plastic with a T-shape, plate-shape, ormushroom shape. The specimen carrier, instead of being made of plastic,can comprise a bio-compatible and inert material (for example,semi-conductor material). The specimen carrier 121 includes a plate-typespecimen receptacle 123 and a carrier pin 124. The inner shape of thepassage openings 111 and the outer shape of the carrier pin 124 arecomplementary for forming a press-fit to one another. Between the edgesof the specimen receptacles 123, gaps 125 are formed. The gaps 125reduce the risk of a mutual contamination between the specimens. Inaddition, they simplify the removable from the cryo-storage elements.With the carrier pins 124, the respective specimen storage 122 issecurely connected.

[0047] On the specimen receptacles 123, the cryo-specimens 130 arearranged, in particular, in the form of frozen liquid drops. The dropsare cell suspensions or also reference specimens, for example, withsamples of cultivation media, solutions of marking dyes, or probespecimens. Probe specimens are reference specimens, which containsubstances that react sensitively to a change of critical environmentalconditions. As probe samples, for example, chemical compounds can beused, which are sensitive to radioactive radiation or unwantedtemperature increases. A control of the probe specimen makes possible amonitoring of the storage condition of the storage substrate in acryo-bank.

[0048] A cover film 114 is arranged over the cryo-specimens 130, whichserves to avoid contamination from the cooling medium or from theenvironment. Typical dimensions of the specimen receptacles 123, forexample, are 0.1 to 3 mm. The entire thickness of the storage substrate100 amounts to approximately 2 mm, for example.

[0049] For the inventive cryo-storage of specimens, a storage substrate100 (without the cover film 114) is first loaded with the cryo-specimensand reference specimens, if necessary. The loading takes place, forexample, with a micro-drop shooting device, such as that described inEP-B 0 804 073. The specimens are shot as micro-drops in a cooled stateof the storage substrate 100, aimed at the specimen receptacles 123,where, upon impingement, they freeze. Likewise, in a deeply cooled stateof the storage substrate 100, writing (for example, a burning) of afirst specimen data takes place in the specimen data storage 122. Afterloading of the substrate, the application of the cover film 114 and theinsertion of the storage substrate in a support under the respectivecooling conditions of the cryo-preservation system that is utilized takeplace.

[0050] In FIG. 2, the removal of specimens from the storage substrate100 is illustrated. According to the present invention, the specimenremoval takes place by separating the respective cryo-storage element120 from the base body 110. The separation takes place with a cuttingdevice 140 in cooperation with a punching device 150. The cutting devicehas a hollow cutting tool 141, whose blade is adapted to the outer shapeof the specimen receptacle 123. The cutting tool, for example, can beformed as a hollow capillary with a ground end. The punching device 150comprises a punch 151, with which the specimen data storage 122 isseparated from the remaining storage medium 112 and the carrier pin 124can be pressed out of the passage opening 111. On the end of the punch151, if necessary, also a cutting tool for improved transection of thestorage medium 112 is provided. The cutting and punching devices 140,150 can be actively or passively cooled for maintaining a predeterminedtemperature of the storage substrate 100.

[0051]FIG. 2 shows a particular advantage of the invention. With thecutting device 140, the specimen 130 is removed with the cryo-storageelement 120, without opening the other specimen deposits. The specimen130 is also connected with the specimen data storage 122 after theremoval. A transfer of the specimen to another storage substrate and/ora measurement device under cryo-conditions or with increased temperaturecan take place. A supplement of specimen data, for example, independence on a measurement result, is provided on the specimen datastorage 122 (data accumulation).

[0052]FIGS. 3 through 7 illustrate inventive storage substrates (forexample, according to FIG. 1) in schematic plan view. A storagesubstrate 100 is formed like a conventional CD and has in the center, inparticular, a passage opening 101 for mounting of the storage substratein the cryo-preservation device and/or a reading/writing system. Thespecimen receptacles 123 of the specimen carrier are formed in thisembodiment as rectangles. They have typical surface dimensions ofapproximately 0.1 to 30 mm². The specimen receptacles 123 are arrangedin groups in sectors 102. Upon removal of specimens with the respectivecryo-storage elements, the base body 110 remains back with the freepassage openings 111.

[0053]FIG. 4 shows a modified embodiment with circular specimenreceptacles 123. On each specimen receptacle 123, a drop with a volumeof some mm³ can be stored. Each drop can contain up to 10⁵ cells. On theentire storage substrate with a diameter of approximately 12 cm,therefore, up to 10⁸ cells can be stored.

[0054]FIG. 5 shows a modified form with circular specimen receptacles123, which, in comparison to FIG. 4, have smaller diameters (forexample, 0.01 to 1 mm). The entire number of the cryo-storage elementson the storage substrate 100 is thereby increased. The variabilityincreases with the specimen removal.

[0055] The base storage 113 illustrated in FIG. 1 can be arrangedselectively according to determined channels in the storage medium. Thisis illustrated in FIG. 6 (ring-shaped storage channels) and 7 (ray-like,aligned storage channels). With the base storage 113, an additionalfragmentation of the storage substrate takes place.

[0056]FIG. 8 illustrates in a cut-away manner a modified form of astorage substrate 200 with a base body 210, which is formed by thespecimen carrier 221 of the cryo-storage element. The specimen carrier221 has the specimen data storage 222 on one side and on the oppositeside, the specimen receptacles 223. The specimen carriers 221 are formedcomponents, for example, made of plastic or a semiconductor material, inwhich the specimen receptacles 223 are formed as recesses. The specimencarriers 221 are connected to one another via breaking points. Thespecimen data storage 222 forms a layer of the storage medium arrangedon the underside of the base body 210. On the upper side of the storagesubstrate, a cover film 214 is provided, with which the specimens 230are covered. The cover film 214 encompasses the base body 210 on itsouter edge with a circulating projection 215.

[0057] The use of the storage substrate 200, in particular, the loadingand the data storage, takes place according to the above-describedprinciples. For data removal, cryo-storage elements 220 are separatedrespectively with a carrier 221 and a specimen data storage 222 with asuitable tool from the storage substrate 220 (for example, broken off,cut off, or the like). Depending on the application, the breaking points224 are designed with a determined geometry, as illustrated in FIG. 9.

[0058] In FIG. 9, the white lines designate the distribution of thebreaking points 224. In the respective framed black regions, thespecimen carriers, in particular, with the specimen receptacles, arearranged. The specimen receptacles can have various geometries withinthe storage substrate 200, for example, toward the interior, they can bemore compact (left drawing part) or narrower (right drawing part).

[0059] A third embodiment of the storage substrate 300 of the inventionis illustrated in FIG. 10. The storage substrate has a disk-shaped basebody 310 in the form of an even, uniform plate. Joints or breakingpoints are not provided in this embodiment. The cryo-storage elements320 form merely one unit in this embodiment, as long as the specimens230 are arranged on the storage substrate 300. As a specimen carrier321, a specimen accommodation layer is provided for each specimen. Thespecimen accommodation layer comprises a plastic material, which has aminimal adhesive strength to the base body 310 (for example, made fromPTFE or rubber). The minimal adhesive strength is provided, inparticular, in low temperature ranges.

[0060] For separating a specimen 330, the specimen is separated with thespecimen accommodation layer from the base body with a suitable tool(for example, detached, planed off, pushed off, or pulled off). Thespecimen data storage 322 remains on the substrate underside.

[0061] The specimens 330 are protected against contamination with acovering 314 in this embodiment. The covering 314 is formed by a cover,which is sealed against the based body 310 via an annular seal 315.

[0062] In FIG. 11, in the upper drawing part in perspective view, acut-away of a storage substrate 400 according to a further embodiment ofthe invention is shown. With this storage substrate, the cryo-storageelements 420 are provided in the form of a pre-perforation or apress-fit in the base body 410. The base body 410 and the cryo-storageelements 420 form a flat plate, on whose underside, the storage medium412 is arranged as a layer. The storage medium 412 (data carrier film)likewise can be pre-perforated and is located on the underside of thebase body 410 with the required planarity for optical reading-in andreading-out of data.

[0063] In the lower drawing part of FIG. 11, the cryo-storage element420 is shown in an enlarged representation. On the top side of the basebody 410, which here forms the specimen carrier 421, a recess isprovided as the specimen receptacle 423. In the specimen receptacle, thecryo-specimen (for example, suspension drop) is arranged. With the coverfilm 414, the specimen 430 is protected against contamination. Also, onthe cover film 414, perforations corresponding to the outer shape of thecryo-storage elements 420 can be provided. According to the invention,generally, the cover or cover film can be formed also as a storagemedium. With removal of the cryo-storage element 420 from the storagesubstrate 400 with a tool formed analogously to the illustration in FIG.2, the specimen 430 is removed with the specimen carrier 420, thespecimen data storage 422, and the cut-out of the cover film 414.

[0064] The storage substrate 400 is fixed in a support and is separatedfrom the base body 410 with the punch 451 (or with the blade 452, ifnecessary) and the cutting device 440. The cutting device 440 isprovided with a movable punch 441, which, after separation of thecryo-storage element 420, presses this out of the cutting device.

[0065]FIG. 12 shows a modification to the embodiment of FIG. 11, by wayof example, of an individual cryo-storage element 420 with the specimencarrier 421 and the specimen receptacle 423. With this form, the cover414 is formed as a two-layered film. A porous layer 415 overlies thespecimen carrier 421 as a cover. Over that, a sealing layer 416 with anextension 417 extending up from the substrate plane is provided. Theextension 417 can be pulled away from the substrate manually or with anappropriate tool. In this manner, the lower layer 415 on the specimencarrier 20 is laid open. This procedure can take place in a deep-freezestate or also in a thawed state. A quick exchange of the liquid in thespecimen receptacle 423 can take place. For example, cryo-protective canbe washed out of the cell suspension.

[0066] The cover 414 can also contain data or markings foridentification of the specimen. According to modified forms, a furtherstructure of the cover 414 formed as additional layers can be provided.

[0067] The principle of the multi-layered cover is also shown in FIG. 13by way of an example of a further embodiment of the invention. There,the storage substrate 500 includes, in turn, a base body 510 withmushroom-shaped projections 511, on which the specimen carrier 521 sits.The specimen carriers 521 are formed components, respectively, with aspecimen receptacle 523 on the upper side and a fixing recess 524 on theunderside. The fixing recesses 524 and the projections 511 work togetherlike push buttons as releasable mechanical connections. On the sideopposite the specimen carriers 521, a data carrier layer as the storagemedium 512 is located, which forms the specimen data storages 522 thatare associated with respective specimen carriers 521. The covering 514takes place according to the double-layer principle, which isillustrated in FIG. 12.

[0068] For removal of a specimen carrier 521 in a frozen state of thestorage substrate 500, a planar or wedge-shaped tool is shoved under thespecimen carrier 521. With the tool, the connection between therespective projection 511 and the fixing recess 524 is loosened. Thespecimen is thereby separated from the storage substrate 500 with thespecimen carrier 521 and parts of the cover 514. Also with thisembodiment, upon separating, the connection to the specimen data storage522 is lost. But, specimen data also can be provided in appropriateparts of the cover 514.

[0069] The embodiment shown in FIGS. 14 through 26 illustrate that thestorage substrate 600 is formed by means of at least one circuit board610, which corresponds to the base body and on which one or morecryo-storage elements 620, like electrical circuits (chips) arearranged. The circuit board 610 supports electrical (conducting paths)or optical (photoconductor fibers) connections 611, which connect,respectively, a receptacle mounting 612 for receiving a cryo-storageelement with an external control device (not shown). The receptaclemounting corresponds essentially to a socket of a conventional circuitmounting, in which the contacts of the cryo-storage element (see, forexample, FIG. 15) are used. On the receptacle mountings 612, additionalcircuits for signal modulation, signal conversion, or detection of thedata signals running from the connection lines 611 or from the opticalpath can be provided.

[0070] The connecting lines 611 can be provided on the top side of thecircuit board 610 with the receptacle mounting 612 (upper drawing partof FIG. 14) or on the opposite side (lower drawing part of FIG. 14). Inthe last case, the receptacle mountings 612 are more tightly arranged.The lower drawing part of FIG. 14 shows further than on the circuitboard 610, also a computing circuit 613 for control of the cryo-storageelements, with the required RAM storage, if necessary, can be provided.

[0071] Each receptacle mounting 612 is equipped for receiving acryo-storage element 620. Each cryo-storage element 620 includes aspecimen carrier 621, analogous to the above-described function, whichis connected with the specimen data storage 622. According to aparticular advantageous embodiment of the invention, the cryo-storageelement is formed by an integrated circuit (for example, storagecomponents) known as such. The circuit contains as the specimen datastorage 622 at least one RAM storage. The cryo-storage element 620 alsocan contain a complete computing circuit, with which the function of thecryo-storage element is executed and by means of which the cryo-storageelement communicates externally. The specimen carrier 621 preferably isformed in or in connection with the plastic covering or encapsulation ofthe integrated circuit.

[0072] The specimen receptacle 623 is, for example, a recess in theplastic over as shown in FIG. 15. With a conventional chip with a sizeof 7·14 mm, the specimen receptacle 423 can have a base surface ofapproximately 4·10 mm with a depth of 1 mm. With these measurements, upto five million cells can be accommodated in the cryo-storage element620.

[0073] On the bottom of the specimen receptacle 623, additional controldevices for manipulation of the specimen sensor and/or display units 624can be provided. The control devices include, if necessary, cooling andheating elements, for example, peltier elements, resistance heatingelements for controlling cooling or heating of the specimen, ormaterials with increased heat capacity for reduction of the heat load ofthe specimen during a chip transport. As a display device, a lightsource can be provided, which, for example, signals a predeterminedstate of the cryo-storage element 620 or the specimen, or which servesas a measured light source for measurement on the specimen 630. Inaddition, the cryo-storage element 620 is provided with a cover 614,which protects the specimen from contamination, evaporation, andsublimation. The cover 614 is a plastic cap, for example, a welded-onfilm, or another layer-type component, which provides a sealed,releasable connection with the specimen carrier 621.

[0074] The specimen carrier 621 serves also as a guide for the contactconnectors 625 of the cryo-storage element. The contact connectors areconnected, in particular, with the specimen data storage 622 and, ifnecessary, to the control and display devices 624.

[0075] In contrast to the diagnosis chips known from cellularbiotechnology, with the cryo-storage element 620, no connection existsbetween the specimen 630 and the specimen data storage 622, the controland/or display devices 624, which is directed to a detection ofelectrical parameters of the cells frozen in the specimen.

[0076] The cover 614 according to FIG. 15 can also be replaced with acryo-container 615, according to FIG. 16. With the cryo-container 615,the top side of the specimen carrier 621 is provided with a sealed screwconnection to a cylindrical container body 617. The cryo-container 615comprises a cold-resistant plastic material. The embodiment of theinvention shown in FIG. 16 has the advantage that the cryo-container 615can also be loaded manually, for example, by means of pipettes.

[0077] Detailed further embodiments of the inventive cryo-storageelements 620 are shown in FIGS. 17 through 26. The cryo-storage element620 according to FIG. 17 includes a specimen carrier 621 and a specimendata storage 622. The specimen data storage 622 is structured like aknown electronic storage chip with contact electrodes 625 and anencapsulation 626, in which a storage circuit and, if necessary, acomputing circuit are arranged. The encapsulation 626 comprisestypically a plastic material.

[0078] The specimen carrier 621 is attached to the top side of theencapsulation 626 or as part of the encapsulation 626. The specimencarrier 621 comprises a plastic frame 627, in which for receivingspecimens, at least one cryo-container 615 is integrated. The plasticframe 627 is an injection molded part with a size corresponding to thesurface of the encapsulation 626, for example. The lateral frame partsare provided with bores, in which the cryo-containers 625 are arranged.

[0079] Each cryo-container 615 forms at least one elongated specimenchamber. The at least one specimen chamber has an elongated form, suchthat the inner cross section is essentially smaller than its length. Asspecimen chambers, for example, hoses, hollow needles, capillaries, orthe like are provided. The internal diameter of a specimen chamber liesin the range of 5 μm to 4 mm, for example. The length can be chosen tobe in the range of 0.5 cm to 10 cm. The quotient of the cross sectionaldiameter and length of a specimen chamber is preferably smaller than{fraction (1/10)}. The allocation of at least one cryo-container 615 ina tube or hose form has the advantage of a fast loading or emptying ofthe specimen chambers, a high ability for miniaturization, and a highfreezing speed.

[0080] With the embodiment of the invention illustrated in FIG. 17,first a cryo-container 615 is formed by a meander-shaped hose placed inthe frame 627 (partially shown). After loading of the cryo-container 615via the hoses (see arrow), the parts of the hose designated with dashedlines are cut off, so that the sections drawn through remain asseparated cryo-containers (for example, 616). All cryo-containers 615are loaded as a unit with a common specimen 630, which advantageously issubdivided into specimen parts. With a mechanical separating device 400,the specimen parts 616 can be separated from the cryo-element 620 in afrozen or thawed state, without affecting the remaining specimen parts.

[0081] On the surface of the encapsulation 626 and/or on the frame 627of the specimen receptacles 623, additional control devices formanipulating of the specimen and/or the sensor and/or the display units624 can be provided. The control devices include, if necessary, coolingand heating elements, for example, peltier elements, resistance heatelements or the like. They serve for the controlled cooling or heatingof the specimen or materials with increased heat capacity for reducingthe heat load of the specimen, for example, during a chip transport. Thesensor and/or display can have a light source, which signals, forexample, a predetermined state of the cryo-storage element 620 or thespecimen or serves as a measured light source for measurement on thespecimen 630.

[0082] In addition, each end of a cryo-container 615 can be providedwith a cover 614, which protects the specimen from contamination,evaporation, and sublimation. The cover 614 is, for example, a plasticcap, a welded-on film or another component, which provides a sealedconnection with the ends of the respective cryo-container 615. If a hoseif provided as a cryo-container, then the cover can be formed also bymeans of a part of the hose itself, which is clamped together on itsends.

[0083] Instead of a through-going and, if needed, cut hose, according toFIG. 17, also a plurality of tube-shaped cryo-containers 615 made from arigid material can be provided as specimen receptacles 623, which arealigned transversely (FIG. 17) or parallel (FIG. 18) to the longitudinalorientation of the cryo-element 620. With the embodiment according toFIG. 18, for example, five cryo-containers 615 are integrated in theencapsulation 626 (enclosed). This can take place by injection of thecryo-container 615 into the encapsulation material or an adhesivebonding. The cryo-containers 615 are formed by hollow needles orcapillaries.

[0084] The loading of a cryo-container 615 takes place, in which on oneend, a low pressure exists and via the opposite input end 617, thecryo-specimen is received. Instead of applying low pressure, also aspecimen receptacle can be provided with the action of capillary forcesin the interior of the cryo-containers 615. In particular, with theembodiment according to FIG. 18, the same or different cryo-specimens631, 632, . . . , can be accommodated in the individual cryo-containers615. The cryo-containers 615 are preferably spaced from one another,such that the feeding ends 617 are aligned to correspond to the formatof a micro- or nanotiter plate.

[0085] According to an alternative embodiment of the inventivecryo-element 620 illustrated in FIG. 19, the cryo-container 615 of thespecimen carrier 622 is formed as a hose, whose diameter in thelongitudinal direction of the hose is changeable. Sections 618 changewith a small diameter and chamber parts 619, in which the hose isconsiderably widened. The cryo-container 615 includes a plurality ofchamber parts 619, which advantageously, can be separated from thecryo-element with a mechanical separating device 400. The cryo-containeris attached on the encapsulation 626 of the specimen storage 622 (forexample, adhered or partially injected). The loading of thecryo-container 615 takes place under formation of a low pressure, whichis absorbed via the input end 617 in the suspended cryo-specimen in thecryo-container 615.

[0086] According to FIG. 20, the cryo-container 615 also can be formedfrom a hose, which has only one chamber part 619, which, as shown, isloaded or emptied via a hose section 618 or alternatively, via multiplehose sections. The chamber part 619 according to FIG. 20 is formed froma film material, which is adhered on the encapsulation 626.

[0087] In contrast to the diagnosis chips known from cellularbiotechnology, with cryo-storage elements 620, no connection existsbetween the specimen 630 on the one hand, and the specimen data storage622 and/or the control, sensor, and/or display devices 624, on the otherhand, which is directed toward a determination of electrical parametersof the cells frozen in the specimen.

[0088] In FIGS. 21 and 22, further details of the loading and thespecimen removal from cryo-elements 620 are illustrated. The loadingtakes place according to FIG. 21, for example, with a loading device,which on one side, has a plurality of specimen reservoirs 710, and onthe other side, has a pressure device 720. The cryo-storage element 620includes a frame-type specimen carrier 621, analogous to the embodimentshown in FIG. 17, on which a plurality of cryo-containers 615 in theform of capillaries or hoses are arranged, whose feeding ends 617project into the specimen reservoir 710. The specimen reservoirs 710 arereservoirs, in which specimens are located after extraction of aspecimen. The pressure device 720 includes a pressure attachment 721,which can be placed in pressure-tight manner on the opposite ends of thecryo-container 615, and a connection line 722, via which allcryo-containers 615 can be pressurized with low pressure. The ends ofthe cryo-container 615 are preferably mounted for common receipt of thepressure attachment 721 on a frame part. They can, for example, openinto the surface of the frame 627. Under the action of the low pressure,specimens 630 are accommodated into the cryo-container 615.

[0089] After the loading of the specimen carriers 621, the cryo-storageelement 620 is separated from the loading device. The input ends 617 areshortened, if necessary, to the frame 627. The cryo-storage element 620is placed on a storage substrate 610 (see FIG. 14), and with this, istransferred in an environment with a reduced temperature, for example, acryo-container in a cryo-bank.

[0090] In FIG. 22, one possibility of specimen removal from acryo-storage element 620 with a meandering-designed cryo-container 615is illustrated. The specimen parts 616 are separated with a mechanicalseparating device 400, for example, a cutting device. This can takeplace advantageously in a state of reduced temperature, so that theremaining specimens can remain unchanged.

[0091] In FIGS. 23 through 26, embodiments of the inventively usedcryo-storage elements 620 are shown, which, in contrast to theabove-described embodiments of the specimen holder 621, is integratednot in an encapsulation of the storage circuit, rather in its substratematerial. FIG. 23 shows, for example, a part of a cryo-storage element620 (without encapsulation and without contact connector). A storagecircuit 800 with a substrate 810 and integrated components 820 are shownin sections. The substrate is a semi-conductor wafer, for example, asare commonly used for manufacturing of integrated circuits. On the upperside of the substrate 810, the components 820 are processed with a knownmethod of semi-conductor technology. On the underside of the substrate810, channel-like specimen chambers 623 are formed as specimenreceptacles, which are closed with a cover layer 820. The specimenchambers 623 have measurements of 400 μm, for example (cross sectionaldimension) and 20 mm (length). The specimen chambers 623 can also beformed in a structured substrate layer, which is arranged on thesubstrate (see FIG. 24). The cover layer 830 comprises a plasticmaterial, glass, or a semiconductor material.

[0092] In addition, multiple levels with specimen chambers 623 can beprovided, as illustrated in FIG. 24. On the substrate 810, a firststructured substrate layer 811 with specimen chambers 623 and a firstcover layer 830 can be provided. On the first cover layer 830, at leastone further structured substrate layer 812 with specimen chambers 623and a further cover layer 831 can be provided. With this embodiment,advantageously, an enlarged specimen volume on the associated specimendata storage is absorbed. The filling or specimen absorption on thecryo-storage elements 620, according to FIGS. 23 and 24, takes place viahose connections, which are integrated in the encapsulation of thespecimen data carrier 622 and are cut off or pinched off after use.

[0093] In FIGS. 25 and 26, various geometries of the channel-shapedspecimen receptacles 623 in the substrates 810 are schematicallyillustrated. For example, meandering or U-shaped channel shapes withvarious cross sectional dimensions can be provided, which, if necessary,are arranged nested in one another.

[0094] The embodiments of the invention shown in FIGS. 14 through 26have a series of advantages. The cryo-storage element is formed by anelectronic chip, which contains an electronic storage that is externallywritable and readable as the specimen data storage. Atemperature-dependent adjustment of a reading/writing head, like that ina CD storage, is not necessary. In the chip, at least one specimenreceptacle corresponding to one or more specimens is located. The chipand/or the receptacle mounting can be equipped with an electronic switchfor controlling additional functional elements, sensors, and/or alarmsystems. The structure shown in FIG. 14 can be made as athree-dimensional multi-level cryo-substrate, in which multiple circuitboards 610 are stacked with a plurality of cryo-storage elements overone another.

[0095] The chip-type storage elements can be removed in a frozen statefrom the circuit board without problems and transferred to other circuitboards, measuring devices, or processing stations, without losing thespecimen data. The cryo-storage elements are electronically addressableexternally.

[0096] In addition, for multiple safeguarding, the cryo-storage elementscan be provided with one or more identifiers, automatically readable orvisually controllable colored markings. The cryo-storage elements,according to FIG. 15, also allow further miniaturization, compared tothe dimensions of common integrated circuits. With miniaturization, anoptimal control is preferred, instead of the electrical contact.

[0097] The at least one circuit board of a storage substrate can beconnected with a computer bus system, with which an individual inquiryand control of the individual cryo-storage elements takes place. Theform of the cryo-storage elements shown in FIG. 15 can be modifieddepending on the application (for example, round or polygonal specimencarriers).

[0098] Important features of the invention are condensed in thefollowing description.

[0099] The inventive storage substrate combines a material receptaclewith a specimen-specific data receptacle. During low-temperaturestorage, the selective removal of materials (cells, cell suspensions)and the reading/storage of data and/or data material are possible.

[0100] The data identification is multiply ensured, in that thespecimens and the specimen data storage are arranged on the same ordirectly adjacent substrate positions. In addition, the storagesubstrate can be dyed, so that based on the color of the cryo-storageelement along and the base body, it can be determined from which storagesubstrate the respective specimen originates.

[0101] The cryo-storage elements can be easily disinfected and reused.Further, they form a protection for the specimens against environmentalconditions with the inventive cryo-preservation.

[0102] It is also possible to provide the base body and the cryo-storageelements of a storage substrate as one unit with a colored or a digitalor analogue identification sample, which allows a clear arrangement ofboth parts at any time. This has advantages for an automated, opticalcontrol (for example, dye and coding detection).

[0103] For the first time, specimen data can be stored in ranges ofkilobytes to megabytes on the inventive cryo-storage. This isparticularly advantageous with the storage of measurement results.

[0104] During the expected useful life of a storage substrate, data canbe supplemented at any time (data accumulation). In this manner, alldata acquired on the specimen or all performed manipulations,measurements, treatments, or the like can be documentedspecimen-specific completely and without interruption.

[0105] Specific treatments of the specimens, in particular, in thechip-type storage elements, can be performed selectively with aprocedure programming and storage. For example, in the frame of acryo-preservation, a predetermined heating, cooling, measuring,controlling, and alarm/display program can operated and be documented ina program data storage. In a frozen state, various temperature ormeasuring programs can run for different cryo-storage elements. Forexample, local thawing can be triggered, in order to perform ameasurement of the specimen. The above-described heating elements can beused with all embodiments of the inventive storage substrate for a localheating of storage media. On the storage media, localized heating canoccur, while the associated specimen remains in a cryo-preserved state.

[0106] According to the present invention, it can be provided that thestorage substrate is operated combined with a mere electronic data bank,in which the specimen data of the storage substrate are stored in amirrored fashion.

[0107] The covering of the specimen receptacles can be partially orcompletely transparent. Through this layer, optical and other measuringmethods are coupled in the specimen receptacles. For example, thespecimens can be shown pictorially. Fluorescent measurements, dielectricmeasurements, and/or ultrasonic representations are possible.

[0108] An inventive cryo-data bank includes a plurality of theabove-described storage substrates, a control device, and a processingdevice for manipulating the storage substrates and for removal ofspecimens.

[0109] The features of the invention disclosed in the foregoingdescription, the claims, and the drawings can be of importance bothindividually as well as in desired combinations for the implementationof the invention in its various embodiments.

1. A method for cryo-preservation, in which a plurality of specimens isarranged on a storage substrate and specimen data, which arecharacteristic for features of a respective one of the specimens, arestored at specific positions.
 2. The method according to claim 1, inwhich the specimen data specimen data are stored on, near, or under theassociated specimen.
 3. The method according to claim 1 or 2, in whichas the specimen data, information for identification of the specimens,information about substance features of the specimens, measurementresults, and/or treatment steps are stored.
 4. The method according toone of the preceding claims, in which the specimen data are read and/orwritten in a cooled state of the storage substrate.
 5. The methodaccording to one of the preceding claims, in which a respective specimenis arranged on a specimen carrier and the associated specimen data arestored in a specimen data storage medium, whereby, respectively, aspecimen carrier and a specimen data storage medium form a compositecomponent that is releasable from the storage substrate for specimenremoval.
 6. The method according to claim 5, in which a specimen isremoved from the substrate storage medium in connection with the storedspecimen data and is transferred onto another substrate or to ameasuring device or a treatment device.
 7. The method according to oneof the preceding claims, in which the specimen data are storedoptically, magnetically, topographically, or electromagnetically.
 8. Themethod according to one of the preceding claims, in which the specimensinclude suspensions of at least one cell, cell component, cellaggregate, and/or tissue.
 9. The method according to claim 8, in whichthe specimens include supplementary reference and probe specimens. 10.The method according to one of the preceding claims, in which, on afrozen, heated or thawed specimen in a cooled state of the remainingstorage substrate, a measurement and/or a treatment takes place andmeasurement results are stored as specimen data.
 11. The methodaccording to one of the preceding claims, in which a control of thestate of at least one specimen on the storage substrate takes place witha computing circuit associated with the specimen.
 12. A storagesubstrate for cryo-preservation of a plurality of specimens, whichcontains a plurality of cryo-storage elements, which are formed,respectively, by means of a composite component from a specimen carrierand a specimen data storage medium.
 13. The storage substrate accordingto claim 12, in which each cryo-storage element is arranged releasablyin a base body of the storage substrate.
 14. The storage substrateaccording to one of claims 12 or 13, in which a moulded component isprovided as the cryo-storage element, which includes the specimencarrier and the specimen data storage medium.
 15. The storage substrateaccording to one of claims 12 or 13, in which an integrated circuit witha storage medium is provided as the cryo-storage element, which isequipped with at least one specimen carrier for respectively receiving aspecimen.
 16. The storage substrate according to claim 15, in which thespecimen carrier is integrated in the structure of the integratedcircuit.
 17. The storage substrate according to claim 16, in which thespecimen carrier has a cryo-container.
 18. The storage substrateaccording to claim 16, in which the specimen carrier has at least onehose-shaped, capillary-shaped, or channel-shaped specimen chamber. 19.The storage substrate according to claim 16, in which the specimencarrier is provided as a channel-shaped specimen receptacle in asubstrate of the integrated circuit.
 20. The storage substrate accordingto claim 16, in which the cryo-storage element contains a computingcircuit, with which the function of the cryo-storage element is managedand by means of which, the cryo-storage element communicates externally.21. A cryo-storage element, which includes a specimen carrier for aspecimen and a data storage medium for storage of specimen data.
 22. Amethod for operating a cryo-bank, in which the specimens arecryo-preserved and/or treated with a method according to one of claims 1through 11.